Team:Ciencias-UNAM/Project/WetLab/Results

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Characterization

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Our experiments focused on the measurements of the expected phenotypes, which in our case, involve the increase or decrease of viability or inhibition. (For details on the protocols, experiment design and photographs pelase refer to the Notebook). The measurement for this in all the experiments listed below, was calculated by measuring the diameter of the inhibition circles in the plates, calculating the area and substracting the area of the inhibition disc.

For the characterization of MarA we used the MarA generator (BBa_K1230004), which is under the Lac promoter. Our first aproach was to test the part with Tetracycline. The results of the experiment are shown in Figure 1. The results show no difference with the differential induction of concentrations of IPTG, but it shows a general interesting behaviour. The cultures transformed with the plasmid show a higher sensitivity to the antibiotic. The control can be observed (in orange) which is a common strain of DH5α.

Due to this unexpected result, we decided to do a second experiment, this time using again Tetracyclin, and also Kanamycin and Ampicilin. The results for the same type of experiment (see Notebook) are shown in Figure 2. This experiment confirmed the results obtained in the first experiment. There was no clear difference between the control and the experimental cultures for Ampicilin, and a slight difference with Kanamycin.

Since Kanamycin showed the most promising results, a third experiment was made using a broader range of this antibiotic. This results are shown in Figure 3.

Again, this figure shows no difference between the IPTG levels, but it shows a very clear difference between the bacteria with the plasmid and the control. This shows an overall increased resistance against Kanamycin.